Agronomic Performance and Seed Inorganic Phosphorus Stability of Low‐Phytate Soybean Line TN09‐239

Phytate (myoinositol‐1,2,3,4,5,6‐hexa‐kisphosphate) in soybean (Glycine max (L.) Merr.) cannot be absorbed by livestock with monogastric digestive systems, and is often excreted in their waste. This can result in agricultural runoff pollution, as well as nutritional deficiencies in poultry (Gallus Gallus domesticus) and swine (Sus domesticus). The enzyme phytase is often applied to break the phytin‐salt bonds and allow for phosphorus (P) absorption, but is an added cost for animal producers. Therefore, we developed a low‐phytate BC₄ ‐ derived line TN09‐239 for comparison of agronomic and seed‐quality traits with the high‐yielding recurrent parent, 5601T. In a replicated, multienvironment field test in Tennessee, TN09‐239 was significantly higher for inorganic P (Pi) (P < 0.001), which is inversely correlated with the seed‐phytate concentration, but significantly lower for yield (P < 0.05) in comparison with the recurrent parent. These findings for increased Pi, but reduced yield, for TN09‐239 in comparison with 5601T were confirmed in the 2010 United States Department of Agriculture (USDA) Uniform Preliminary V Soybean Test grown in 10 southern US environments. 5601T and TN09‐239 differed in Pi stability across southern environments, with linear regression showing 5601T having stability across environments, while TN09‐239 showed variations based on the environments. However, in each environment TN09‐239 displayed nearly a 10‐fold increase in Pi compared to 5601T. Although the low‐phytate trait and much of the recurrent parent genome along with 85–90% of the yield have been captured, further backcrossing could help recover the remaining seed yield of 5601T and other traits desirable for southern US producers.     

Generation and Characterization of a Soybean Line with a Vernonia galamensis Diacylglycerol Acyltransferase-1 Gene and a myo-Inositol 1-Phosphate Synthase Knockout Mutation

Soybean (Glycine max) meal is an important protein source. Soybean meal with lower phytate and oligosaccharides improves meal quality. A single recessive mutation in soybean myo-inositol 1-phosphate synthase (Gm-lpa-TW75-1) confers a seed phenotype with low phytate and increased inorganic phosphate. The mutant was crossed with high oil lines expressing a diacylglycerol acyltransferase1 (DGAT) gene from Vernonia galamensis (VgD). Gm-lpa-TW75-1 X VgD, designated GV, has 21%, and 22% oil and 41% and 43% protein from field and greenhouse seed production, respectively. No significant differences were found in mineral concentrations except for Fe which was 229 μg/g dry mass for GV followed by 174.3 for VgD and 162 for Gm-lpa-TW75-1. Phosphate (Pi) is higher in Gm-lpa-TW75-1 as expected at 5 mg/g, followed by GV at 1.6 mg/g whereas Jack, VgD, and Taiwan75 have about 0.3 mg/g. The Gm-lpa-TW75-1 line has the lowest phytate concentration at 1.4 mg/g followed by GV with 1.8 mg/g compared to Taiwan75, VgD, and Jack with 2.5 mg/g. This work describes a high oil and protein soybean line, GV, with increased Pi and lower phytate which will increase the nutritional value for human and animal feed.     

Does the Pi strip method allow assessment of the available soil P?: Comparison against the reference isotope method

Experiments have been carried out in acid soils developed under tropical climates with and without phosphate rock (PR) addition to assess the ability of the Pi strip method to extract available soil P compared with the Isotopic Exchange Kinetics method (IEK). In the Pi strip method strips of filter paper, previously impregnated with iron hydroxide acting as a sink for phosphate ions from soil components, are added to an aqueous soil suspension. The extracted phosphate ions are eluted in diluted sulfuric acid and quantified by a colorimetric method. Available soil P, defined as the amount of phosphate ions that can move from the soil particles to soil solution, is described by three factors: an intensity factor, a quantity factor and a capacity factor. These three factors were determined by the IEK-method. Following the addition of carrier-free ³²PO₄-ions to soil, the ability of the Pi strip to extract available soil P was assessed: (i) by comparing the quantity of instantaneously exchangeable P (E₁) to the quantity extracted with the Pi strip; (ii) by determining the fraction of ³²P extracted with the Pi strip, and (iii) by comparing the specific activity (SA) of P present as phosphate ions extracted by the Pi strip to the specific activity of P in the soil solution. It was observed that (i) E₁ and the amount extracted with the Pi strip are highly correlated, (ii) the recovery of ³²P extracted by the Pi strip varies between 17 and 66%, and (iii) the specific activity of P extracted by the Pi strip is of the same order of magnitude as that of P in the soil solution. In acid soils low in available P, part of the P in aqueous KCl-extracts is presumably not only present as free phosphate ions but also occluded in the form of a soluble complex, whose isotopic exchangeability is significantly lower than that of phosphate ions transferred to the Pi strip. It is concluded from the results that the Pi strip method can be recommended in routine analysis for the determination of the quantity factor. However, this method cannot provide intensity or capacity factors and therefore needs to be complemented by the IEK-method for full characterization of the available soil P status.     

Five‐Membered Cyclitol Phosphate Formation by a myo‐Inositol Phosphate Synthase Orthologue in the Biosynthesis of the Carbocyclic Nucleoside Antibiotic Aristeromycin

Aristeromycin is a unique carbocyclic nucleoside antibiotic produced by Streptomyces citricolor. In order to elucidate its intriguing carbocyclic formation, we used a genome‐mining approach to identify the responsible enzyme. In silico screening with known cyclitol synthases involved in primary metabolism, such as myo‐inositol‐1‐phosphate synthase (MIPS) and dehydroqunate synthase (DHQS), identified a unique MIPS orthologue (Ari2) encoded in the genome of S. citricolor. Heterologous expression of the gene cluster containing ari2 with a cosmid vector in Streptomyces albus resulted in the production of aristeromycin, thus indicating that the cloned DNA region (37.5 kb) with 33 open reading frames contains its biosynthetic gene cluster. We verified that Ari2 catalyzes the formation of a novel five‐membered cyclitol phosphate from d ‐fructose 6‐phosphate (F6P) with NAD⁺ as a cofactor. This provides insight into cyclitol phosphate synthase as a member of the MIPS family of enzymes. A biosynthetic pathway to aristeromycin is proposed based on bioinformatics analysis of the gene cluster.     

硅钼兰比色法测定锅炉水中二氧化硅的优化

通过探讨分光光度法测定水中常量二氧化硅的方法,在盐酸浓度为0.12 mol/L的酸度条件下,水样中活性硅与钼酸铵生成为硅钼黄,酒石酸掩蔽磷酸根等干扰物质,用抗坏血酸还原生成硅钼蓝,生成硅钼蓝络合物的颜色深度与水样中硅浓度成正比,符合郞伯比尔定律。本方法适用于SiO_2含量较高的水样中SiO_2的测定,如原水、循环冷却水和炉水等。此方法操作简单,准确度高,具有推广价值。     

Characterization of polar and nonpolar seed lipid classes from highly saturated fatty acid sunflower mutants

The seed lipids from five sunflower mutants, two with high palmitic acid contents, one of them in high oleic background, and three with high stearic acid contents, have been characterized. All lipid classes of these mutant seeds have increased saturated fatty acid content although triacylglycerols had the highest levels. The increase in saturated fatty acids was mainly at the expense of oleic acid while linoleic acid levels remained unchanged. No difference between mutants and standard sunflower lines used as controls was found in minor fatty acids: linolenic, arachidic, and behenic. In the high-palmitic mutants palmitoleic acid (16∶1n−7) and some palmitolinoleic acid (16∶2n−7, 16∶2n−4) also appeared. Phosphatidylinositol, the lipid with the highest palmitic acid content in controls, also had the highest content of palmitic or stearic acids, depending on the mutant type, suggesting that saturated fatty acids are needed for its physiological function. Positional analysis showed that mutant oils have very low content of saturated fatty acids in the sn-2 position of triacylglycerols, between the content of olive oil and cocoa butter.     

烷基磷酸酯中有效磷的测定

用抗坏血酸-磷钼蓝分光光度法测定烷基磷酸酯中的有效磷。有效磷=总磷-无机磷。考察了检测波长、显色剂用量、硫酸加入量及显色时间等测定条件对测定结果的影响。方法最佳测定条件为:检测波长820nm;硫酸(9.58 mol.L-1)、钼酸铵(60 g.L-1)、抗坏血酸(100 g.L-1)的加入量分别为2.00、2.00、1.00 mL,70℃水浴条件下显色20 min。在该条件下,测定吸光度值与质量浓度呈线性关系,总磷测定回归方程为:A=0.1924ρ+0.0134,相关系数r=0.999 5;无机磷用正戊醇萃取法测定,回归方程为:A=0.1846ρ+0.0094,相关系数r=0.999 1。方法回收率为91%~97%,RSD小于3.4%。该方法操作简单、计算方便,有望应用于磷酸酯生产的质量控制。     

Isolation of chemically induced mutants in borage (Borago officinalis L.)

γ-Linolenic acid (GLA, 18:3 Δ6,9,12) has been reported to be helpful in the treatment of a wide range of disorders. Borage (Borago officinalis L.) is an annual plant of renewed interest because the seeds are an important source of GLA. The failure to retain mature seeds until harvest limits the total seed and GLA yield per plant and is the major limiting factor for the commercial production of borage. In the course of a mutagenesis program, an agronomically good line of white-flowered borage (RG-001) was treated with ethyl methane sulfonate. As a result of this program, several types of mutants were identified in the M2 generation of plants: a chlorotic mutant (type A); a mutant with increased number of sepals, petals, and ovules but reduced fertility (type B); and mutants with closed flowers (type C1) or partially opened flowers (type C2) that had increased seed retention. The type C mutants are the first reported borage plants with a nonshattering habit. After crossing type B plants with normal plants, a new mutant (type B1) was obtained with higher fertility and higher seed production per flower than those from normal plants. These mutants could be used to develop borage lines that would be superior to those currently available as a source of GLA.     

磷酸中五氧化二磷的快速测定

磷钼酸喹啉重量法测定P2O5需4 h,故实验研究采用钒钼酸铵分光光度法测定P2O5。提取磷酸试液中正磷酸根离子,在酸性介质中与钼酸铵及偏钒酸盐反应,生成稳定黄色配位化合物,于波长420 nm处,测其吸光度,即可求出五氧化二磷含量,方法快速,准确度和精密度符合中控要求。     

Phytic acid in soybeans

Phytic acid, the hexaphosphate of myo-inositol, is the most important phosphate reserve compound in many plant seeds, but many of its salts are poorly digested by animals. It can form complexes with seed proteins, some of which sequester metal ions, making them unavailable for the animal organism. Soya protein isolates may be higher in phytate content than the soya flour from which they are obtained. Zinc is the mineral of most concern because its bioavailability from some soya products is quite low and because of its marginal levels in some human diets. The availability of iron from soya flour and soya isolates is higher than that from some other plant foods with lower phytate contents. Processes for removing the larger part of the tightly bound phytates from soya protein isolates are described.     

Phosphorus seed coating increases phosphorus uptake,early growth and yield of pearl millet (Pennisetum glaucum (L.) R. Br.) grown on an acid sandy soil in Niger,West Africa

In pot and field experiments conducted in 1990 and 1991 on an acid sandy, phosphorus (P) deficient soil in Niger, West Africa, the effect of seed coating on seedling emergence, early growth and grain yield of pearl millet (Pennisetum glaucum (L.) R. Br.) was studied. Seeds of pearl millet were coated with different rates (0; 0.5; 1.0; 2.0; 5.0; 10.0 mg P seed^–1) and types of P fertilizers (single superphosphate, ammonium dihydrogen phosphate; monocalcium phosphate, sodium dihydrogen phosphate and sodium triphosphate). Seedling emergence was generally reduced at coating rates higher than 0.5 mg P seed^–1 and prevented with single superphosphate and sodium triphosphate at rates higher than 5 mg P seed^–1. No correlation was found between the pH and osmomolity of the coatings and final emergence of millet seedlings. The most favourable effect on plant growth and P content was achieved with ammonium dihydrogen phosphate (AHP) as seed coating. This was attributed to the enhancement effect of ammonium on P uptake. Compared to the untreated control dry matter production at 20 days after planting (DAP) was increased by 280%, P content per plant by 330%, total biomass at maturity by 30% and grain yield by 45%. Although seed coating with AHP may be harmful to seedlings emergence, it represents a suitable method to enhance early growth and increase yield of pearl millet.     

Hydrolyzed collagen‐grafted‐poly[(acrylic acid)‐co‐(methacrylic acid)] hydrogel for drug delivery

Hydrolyzed collagen‐grafted‐poly[(acrylic acid)‐co‐(methacrylic acid)] hydrogels were synthesized by solution polymerization and confirmed by infrared spectroscopy. From sequential univariate analysis, the optimal molar ratio of acrylic acid: methacrylic acid was 92:8 in the presence of N,N′‐methylenebisacrylamide, ammonium persulfate, and N,N,N′,N′‐tetramethylethylenediamine at 0.12, 0.015, and 0.2% mol of the monomers, respectively. The water absorbency of this hydrogel was both pH‐ and temperature‐dependent, but was higher in nonbuffered water than in boric acid/citric acid/phosphate buffer under the same conditions. The optimal hydrogel could swiftly swell and deswell in neutral and acid solutions, respectively. Its potential application in drug delivery was examined using insulin and methylene blue as model payload drugs. Loading in a 50% (v v^?1) ethanol solution gave a higher insulin loading level than in the buffer water. Insulin and methylene blue were both released at pH 6.8 but not at pH 1.2, but followed first order kinetics and the Higuchi equation, respectively. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018 , 135, 45654.     

铋磷钼蓝分光光度法测定硅铁中磷的研究

研究用抗坏血酸还原,磷与钼酸铵、硝酸铋形成蓝色的显色反应。分析了酸度、显色时间的影响,确定了最佳测定条件。结果表明,加标回收率达97．88％。此方法简单、快速且稳定,能获得满意的分析结果。     

Nonhypothesis Analysis of a Mutagenic Soybean (Glycine max [L.]) Population for Protein and Fatty‐Acid Composition

Soybean is a major source of oil for food, feed, and biofuel production. Mutagenesis is a tool for creating unique traits useful in breeding programs. The aim of this study is to use nonhypothesis statistical testing methods to make decisions about a mutagenic population. To this end, a total of 1037 mutation lines and 28 wild‐type lines were analyzed for fatty‐acid composition and protein content. Principal component analysis (PCA) was used to analyze the fatty acid profile, multivariate analysis of variance (MANOVA) to build a selection model for seed weight per plant and weight per 10 seeds, and clustering in conjunction with power analysis to determine the minimum number of individuals needed to create a MANOVA selection model for the oil to protein content. Five of the 35 possible entries were identified by PCA analysis for stearic acid and four of 16 possible entries for oleic acid. Interestingly, most of the selected mutants were validated genetically. In fact, selected mutants with high seed stearic acid or high seed oleic acid contents were verified to carry mutations on GmFAD2‐1A, GmFAD2‐1B, and GmSACPD‐C genes. This shows a promising method of identifying smaller portion of the population to screen for desired mutations.     

New FATB thioesterases from a high‐laurate Cuphea species: Functional and complementation analyses

The New World genus Cuphea is a rich source of genes encoding enzymes specialized for seed‐specific synthesis of short‐ and medium‐chain fatty acids (FA). We isolated and characterized two new acyl‐acyl carrier protein (ACP) thioesterase (TE) cDNA, CcFATB1 and CcFATB2, from developing seeds of C. calophylla var. mesostemon. Cuphea calophylla is one of several species in the genus specializing in the synthesis of lauric acid (12:0). Phylogenetic clustering classified CcFATB1 and CcFATB2 as medium‐ and long‐chain FATB TE, respectively. Gene expression analysis indicated that CcFATB1 was seed specific and CcFATB2 was ubiquitous. Overexpression of CcFATB1 in developing seeds of Arabidopsis partially redirected FA synthesis toward medium‐chain FA (12:0, 14:0, and 16:0). Laurate synthesis was preferentially increased by co‐expressing CcFATB1 and CwKAS A1, a medium‐chain‐specific ketoacyl‐ACP synthase (KAS) from another 12:0‐rich species, C. wrightii. Overexpression of CcFATB1 cDNA complemented the low‐palmitate (16:0) but not the low‐stearate (18:0) phenotype in leaves and seeds of an Arabidopsis fatb‐2 mutant, whereas CcFATB2 restored both long‐chain FA. The investigated Cuphea species provide a diverse source of seed‐specific FATB and KAS genes for combinatorial manipulation of medium‐chain FA content in oilseed plants.     

Physical‐chemical characteristics and oxidative stability of oil obtained from lyophilized raspberry seed

Fresh raspberry (Rubus idaeus), cultivar Willamette, was freeze‐dried (lyophilization). A byproduct of lyophilization is “fine dust” of raspberry consisting of finely ground raspberry fruit body and seed. The seeds were separated. The seed oil was isolated and its physical and chemical characteristics were determined. Parameters that characterize the seed and quality of the oil were examined, including fatty acid composition, oxidative stability under different storage conditions, and radical‐scavenging activity. The fatty acid composition was determined by GC/FID and the contents of the dominant fatty acids were found as: oleic 16.92%, linoleic 54.95%, and α‐linolenic acid 23.97%. The oxidative stability of the oil was poor. The induction period by Rancimat test at 100 °C was 5.2 h. The radical‐scavenging activity is similar to that of resveratrol [1,3‐benzenediol 5‐(1^E‐2‐4‐hydroxy‐phenyl‐ethyl)]. Although this product is used in the candy industry, it would be far more useful if raspberry oil of satisfactory quality could be extracted. This paper demonstrates that sifted lyophilized seeds can be used for the extraction of oils. This process allows for maximal usage of the byproducts, reduces losses and it increases the development of new products.     

Enzyme‐assisted aqueous extraction of oil and protein from canola (Brassica napus L.) seeds

The emphasis of this study was to investigate the effect of enzymes on aqueous extraction of canola (Brassica napus L.) seed oil and protein. Four enzymes, Protex 7L, Multifect Pectinase FE, Multifect CX 13L, and Natuzyme, were tested for their effectiveness in releasing oil and protein during aqueous extraction. The enzyme‐extracted oil content of canola seeds (22.2–26.0%) was found to be significantly (p <0.05) higher than that of the control (without enzyme) (16.48%). An appreciable amount of protein (3.5–5.9%) originally present in the seed was extracted into the aqueous and creamy phases during aqueous extraction of oil. The physicochemical properties of oils extracted from canola seed by conventional solvent extraction, and aqueous extraction, with or without enzyme addition were compared. Significant (p <0.05) differences were observed in free fatty acid content, specific extinctions at 232 and 270 nm, peroxide value, color (1‐inch cell) and concentration of tocopherols (α, γ, and δ). However, no significant variation (p <0.05) was observed in iodine value, refractive index (40 °C), density (24 °C), saponification value, unsaponifiable matter and fatty acid composition. A better oil quality was obtained with aqueous extraction (with and without enzyme) than with solvent extraction. While the enzymes enhanced the oil extraction, the oil yield was still significantly (p <0.05) lower than that obtained by solvent (hexane) extraction.     

Proximate Composition and Fatty Acid Profile of <Emphasis Type="Italic">Pongamia pinnata</Emphasis>, a Potential Biodiesel Crop

The chemical characteristics of Pongamia pinnata seeds, focussing on proximate composition and the fatty acid profile of its oil, are presented. The proximate composition of P. pinnata seeds was: 3.8% ash, 9.7% sugar, 7.07% protein, 24% oil, 10.7% free amino acids, and 0.24% free fatty acids. The oil was extracted from seeds by use of different solvents and the highest yield (29%) was obtained by use of n-hexane. Monounsaturated and polyunsaturated fatty acids accounted for 63.3 and 22.9%, respectively, of the seed oil. Oleic acid was the major fatty acid but a substantial amount of erucic acid was also detected; this was not reported in previous studies. The level of erucic acid and the presence of toxic flavonoids, for example karanjin, pongapin, and pongaglabrin, render the oil inedible according to WHO recommendations. However, low levels of saturated and polyunsaturated fatty acids with desirable cetane number and iodine value suggest potential for application as a biodiesel fuel.     

草酸-酸亚硅钼蓝光度法检测钢铁及合金中硅的含量

试样用稀酸溶解,在微酸性溶液中,硅酸与钼酸铵生成硅钼杂多酸,在草酸存在下,用硫酸亚铁铵还原成硅钼蓝,测其吸光度.与钼酸盐形成杂多酸的硅必须以正硅酸状态存在,所以在试样溶解过程中必须避免硅酸的聚合.在溶液中硅酸较易于聚合,其聚合程度与溶液的酸度、溶解时加热的程度和时间以及最终溶液中正硅酸离子的浓度有关     

Nondestructive determination of fatty acid composition of husked sunflower (Helianthus annua L.) seeds by near-infrared spectroscopy

Determination of the fatty acid composition of sunflower (Helianthus annua L.) seeds by near-infrared (NIR) spectroscopy was examined. Sunflower seeds were husked (removed from their hulls by a husking machine or manually with a knife). NIR spectra of these seeds were scanned from 1100 to 2500 nm at 2-nm intervals in a whole-grain cell with a wideangle moving drawer for machine-husked seeds or in a single-grain cup for a manually husked single-grain seed. The extracted oils from machine-husked seeds also were scanned by sandwiching them between a pair of slide glasses to create a thin layer and by placing them on a syrup cup. For extracted oil, the absorption band around 1720 nm filled out to the shorter wavelength region in the NIR second-derivative spectra as the percentage of the linoleic acid moiety increased, because linoleic acid absorbs in this region. On the other hand, for husked seeds and for a single-grain seed, as the percentage of linoleic acid increased, the trough at 1724 nm where oleic and saturated acids absorb decreased in the second-derivative NIR spectra. Determination of the fatty acid composition of sunflower seeds could be carried out successfully according to the NIR spectral pattern for both extracted oil (r=−0.989) and kernel seed (r=−0.993). This is important, especially for a manually husked single-grain seed (r=−0.971), because it can still be germinated after such nondestructive analysis.